PhD Scientific Days 2024

Pharmaceutical Sciences and Health Technologies II.

Scalibity of spheroid-derived extracellular vesicles production in stirred system

Előadó neve

Ms. Dauphin, Thibaud

Előadó munkahelye

Oniris

Előadó telefonszáma

+33749875281

Előadó e-mail címe

thibaud.dauphin@oniris-nantes.fr

Az előadás címe

Scalibity of spheroid-derived extracellular vesicles production in stirred system

Szerző(k) neve és munkahelye

Thibaud Dauphin¹, Laurence de Beaurepaire¹, Apolline Salama¹, Clémentine Claire¹, Quentin Pruvost¹, Karine Haurogné¹, Sophie Sourice¹, Aurélien Dupont², Jean-Marie Bach¹, Eric Olmos³, Steffi Bosch¹, Blandine Lieubeau¹, Mathilde Mosser¹

1: Oniris, INRAE, IECM, Nantes, France
2: University of Rennes 1, CNRS, Mric, Rennes, France
3: University of Lorraine, CNRS, LRGP, Nancy, France

Bemutatás módja

Szóbeli

Szekció

Pharmaceutical Sciences and Health Technologies II.

Language of the presentation

English

Preferred session

Pharmaceutical Sciences and Health Technologies

Összefoglaló szövege

Introduction: Extracellular vesicles (EV)-based therapies have gained widespread interest, but efforts remain to ensure standardisation and high-scale production. Implementing processes in stirred-tank bioreactors (STR) is crucial to finely control the cell environment and scaling-up production, but it remains a significant challenge for adherent cells. Here, we investigate the impact of culturing a pancreatic cell line as spheroids in stirred systems on cell fate and EV release. In order to fix the agitation rate for large-scale production, we assessed whether the power input (P/V) or impeller tip speed should be keep constant.
Aims: We aim to establish a standardised process for spheroid-derived EV production and evaluate the influence of culture mode on EV properties.
Methods: 1.4E7 cells were cultured as spheroids in spinner flasks (SpF) or ambr®250 STR. After 24h of culture in serum-free medium, EV were isolated by tangential flow filtration and size exclusion chromatography. EV size and concentration were determined by nanoparticle tracking analysis of tetraspanin-labelled particles. Protein expression was detected by western blot. EV immune properties were assessed through mixed lymphocytes reaction (MLR).
Results: Culturing 1.4E7 cells in suspension reduced their specific growth rate but allowed the formation of homogeneous and viable spheroids. Spheroid formation is dependent of the P/V, and maintaining this criterion constant across scales proved to be the optimal scale-up strategy. However, the process transfer to STR altered spheroid fate due to impeller design and its impact on flow regime. Compared to a monolayer process, EV yield decreased (2-fold) in SpF but increased in STR (2-fold), and EV exhibited distinct immune responses in MLR.
Conclusion: As EV are the mirror of their parental cells, it is crucial to understand and master the culture environment and to fine tune each process parameter during set-up and scale-up to warrant EV final quality and quantity attributes. We present a comprehensive framework addressing key challenges encountered in developing scalable EV-based therapy production from anchorage-dependent cells.
Funding: it is supported by the French National Research Agency (ANR), the Ministry of Agriculture and Food Sovereignty, and the "Pays de la Loire" region.

University

Nantes Université

Supervisor

Blandine Lieubeau

Publication of my abstract

I give consent to the publication of my abstract on the website of the congress.

Kind

Szabad

Status

elfogadva

Accepted presentation method

szóbeli

Előadás fájl jóváhagyás

nem rendelkezett róla

Előadó

8251

Start

11:00

End

11:10